EP3353549A1 - Biocapteur et procédé de détection de germes - Google Patents

Biocapteur et procédé de détection de germes

Info

Publication number
EP3353549A1
EP3353549A1 EP16767176.7A EP16767176A EP3353549A1 EP 3353549 A1 EP3353549 A1 EP 3353549A1 EP 16767176 A EP16767176 A EP 16767176A EP 3353549 A1 EP3353549 A1 EP 3353549A1
Authority
EP
European Patent Office
Prior art keywords
test strip
biosensor
lateral flow
germ
flow test
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP16767176.7A
Other languages
German (de)
English (en)
Inventor
Petra Neff
Katrin Luckert
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Robert Bosch GmbH
Original Assignee
Robert Bosch GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Robert Bosch GmbH filed Critical Robert Bosch GmbH
Publication of EP3353549A1 publication Critical patent/EP3353549A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54386—Analytical elements
    • G01N33/54387—Immunochromatographic test strips
    • G01N33/54388—Immunochromatographic test strips based on lateral flow
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses

Definitions

  • Biosensor and method for detection of germs The present invention relates to a biosensor in the form of a lateral flow
  • Test strip and a method for the detection of germs.
  • Molds and above all mold spores and toxins secreted by mold fungi are not to be underestimated as pathogens of infections and chronic diseases, for example inflammatory lung diseases. Furthermore, molds and mold spores are potential allergens that can lead to the triggering of allergic complaints. Mold infestation or, in general, microbial contamination can occur, in particular, within various domestic appliances, for example in washing machines or coffee machines. today
  • washing powder compartment can come. Even modern coffee machines, especially fully automatic machines, are often exposed to microbial contamination, as compared to conventional filter coffee machines for cleaning difficult and cumbersome achievable areas are available due to the complex structure.
  • the different moist locations in a modern coffee machine eg Water tank, hoses, pomace, grinder
  • a modern coffee machine eg Water tank, hoses, pomace, grinder
  • Mold fungi may have very good growth conditions.
  • Washing programs or cleaning programs are carried out, which should reduce a germ load. Since such cleaning measures in practice, however, may not be carried out regularly and an occurring bacterial load is not noticed, it can come by the unrecognized bacterial load to health problems for the user.
  • a hand-held measuring device for the detection of hidden mold damage is known. This can be a mold in indoor areas, such. B. on walls can be detected.
  • the device includes, among other things, a collection unit for receiving an interior sample and an immunological test unit.
  • German Patent Application DE 196 04 195 A1 discloses an analysis method in which antibodies or antigens are aspirated from the room air and subsequently detected by an immune reaction.
  • the invention relates to a biosensor for the immunological detection of germs, wherein the biosensor according to the invention is designed as a lateral flow test strip with germ-specific antibodies.
  • the mobility characteristics of the antibodies change as the antibodies complex with the germ to be detected. This change in mobility characteristics will be
  • biosensor used according to the invention to detect germs with the lateral flow test strips in a particularly simple and user-friendly manner.
  • Preferred embodiments of the biosensor are the subject of the dependent claims.
  • the biosensor according to the invention which is based on an immunological detection, is very sensitive, so that it is also possible to detect germs, which under certain circumstances occur in relatively low concentrations. Furthermore, the selectivity is due to the use of highly specific antibodies, which are highly specific antibodies, and many others.
  • Biosensors a possibly beginning incipient germs are detected at an early stage, so that appropriate measures, such as the cleaning of a germ-loaded household appliance,
  • the biosensor is designed in the form of a lateral flow test strip.
  • a lateral flow test is a detection method based on a combination of thin layer chromatography and immunostaining.
  • the sample optionally together with bound antibodies, migrates over the length of the test strip due to capillary forces (thin-layer chromatography). Since the
  • the mobility properties of the antibodies change when the antibodies complex with the germ to be detected, as a result of which they depend on the presence of the germs to be detected
  • At least one detection reagent is bound to the germ-specific antibodies.
  • the detection reagent is directly or directly bound to the germ-specific antibody.
  • the detection reagents can be covalently immobilized (coupled) via amino acid groups.
  • COOH / NH2 can be coupled.
  • other reactive groups of the antibody e.g. Cysteine / SH groups, when using linker systems for chemical bonding with the
  • the detection reagent is metal particles or dye molecules.
  • an optical read-out system can be realized in a very user-friendly manner, wherein it can be recognized by means of a color reaction, which can be evaluated, for example, with the naked eye, whether there is a germ load or not. In particular, quantitative detection is generally inaccurate or even impossible to visualize with the eye.
  • optical devices may also be used in other embodiments, eg cameras, which in particular also permit a quantitative or semiquantitative evaluation on the basis of a measurement of color gradations or intensity graduations.
  • the evaluation can be software-based, for example, a comparison with eg
  • the stored reference values can be made.
  • the camera of a mobile phone can be used as an optical device.
  • the biosensor is provided for the detection of molds and / or mold spores and / or bacteria. These mentioned germs are relevant in the first place for a possible germ load of household appliances. A bacterial infestation or a mold infestation or infestation with mold spores occurs especially in permanently moist places in household appliances, for example in water tanks or hoses, or in places where moist material stored for a long time, for example in a pomace container.
  • the biosensor can be directed to the detection of a particular germ. Depending on the device and application case, it is preferably determined in advance which germ could be mainly responsible for a load.
  • the germ-specific antibodies are directed against surface molecules of the germ to be detected.
  • the antibody may preferably be directed against an epitope on the surface of the germs. This has the advantage that no further sample preparation, z. B. no disruption of cells or the like, must be done.
  • the germ-specific antibodies can bind directly to the germs to be detected, so that the immunological detection can be done in a very simple manner.
  • Detection system is therefore aligned the biosensor on the detection of certain germs.
  • test strips that are set up for the detection of molds and test strips that are set up for the detection of specific bacteria can be provided.
  • several different test strips can be used in parallel or sequentially for the detection of certain germs.
  • test strip itself can also be designed so that it is set up for the parallel detection of different germs.
  • the lateral flow test strip is designed as a disposable article. Following use of the biosensor and bacterial loading test, the test strip may then be discarded, with the germs collected and analyzed on the test strip, together with the test strip, disposed of directly in the household waste. The accumulated on the test strip germs are usually not dangerous goods. If the lateral flow test strip as
  • Disposable article is designed, it has the further particular advantage that such an article is generally very inexpensive to produce.
  • the lateral flow test strip comprises a sample-receiving pad and / or a capillary and / or at least one detection field.
  • the sample-receiving pad which is the Sample collection and also conjugation with the assay components (antibodies) is preferably adjusted to the sample in terms of pore size and binding properties.
  • the sample-receiving pad may be formed, for example, from nitrocellulose and be realized for example in the form of a membrane. Furthermore, for example, cellulose or
  • test strip itself may for example be based on a strip of plastic as a carrier.
  • capillary is meant an area of the test strip that can be traversed by liquids due to capillary forces. Generally suitable for this
  • liquid-permeable materials for example nitrocellulose
  • Polyvinylidene fluoride (charge-modified) nylon or polyethersulfones.
  • the lateral flow test strip preferably comprises a control field, so that the user can, for example, by direct comparison of the
  • Check field with the detection field can check whether a positive or a negative test result.
  • the germ-specific antibodies may for example be presented in dried form on the test strip or be added from a separate solution.
  • the capillary is designed so that after a complex formation of the antibody with the optionally
  • test strip may contain one or more buffers, for example recording buffer and / or detection buffer and / or running buffer.
  • the lateral flow test strip may be assigned a buffer cartridge, from which any buffer required is released.
  • the sample is applied to the sample receiving pad in liquid or solid form.
  • a required buffer for example a running buffer, is released from the buffer cartridge and applied, for example, in the region of the sample-receiving pad, so that the sample can be transported by capillary forces.
  • the sample is applied in liquid form (for example, a water sample from a water tank), it may also be sufficient for the sample to be applied
  • Liquid is taken from the sample. In this case, a separate run buffer is not required. If a sample is tested in dry form (eg coffee residues from the grinder), manual or automatic addition of buffer is required.
  • a pre-stored in a buffer cartridge buffer can be released manually or automatically and applied directly or via a channel, for example by means of capillary forces on the sample receiving pad.
  • the buffer cartridge (for example a plastic container) can be designed, for example, so that the buffer is released by kinking.
  • a cell disruption reagent e.g. a detergent
  • a buffer solution e.g. a buffer solution
  • the antibodies By means of thin-layer chromatographic separation, the antibodies, if appropriate together with the germs to be detected in complex form, reach the detection field and / or a control field of the lateral flow test strip.
  • the test result is preferably by a colored reaction or a
  • Color change visible to the user Colored readout systems are particularly preferred for this, so that there is no need for further aids for the evaluation of the test strip.
  • optical devices e.g. the camera of a mobile phone or otherwise, be used for the evaluation.
  • biosensor according to the invention for an immunological detection of germs in one
  • the biosensor according to the invention thus allows the user a independent inspection of potentially infested areas, for example in a washing machine or coffee machine. After the test has been carried out, the user receives a qualitative or semiquantitative or possibly quantitative result, so that suitable cleaning measures can be taken if necessary for a positive test result.
  • the shape and configuration of the biosensor or the lateral flow test strip can be adapted to the intended sampling location in the household appliance.
  • the lateral flow test strip can be configured in a conventional rod shape.
  • the test strip is adjusted accordingly, for example by a
  • test strip can be inserted, for example, in a hose or the like. You can also continue
  • Sampling sticks or the like may be provided, with the aid of, for example, a swab sample is taken in the device, which is then applied to the test strip.
  • the invention further relates to the use of a biosensor in the form of a
  • Lateral flow test strip for the immunological detection of germs in a household appliance, in particular in a washing machine or in a
  • Coffee machine With regard to further features of the lateral flow test strip and in particular with regard to the use for detecting germs in a domestic appliance, reference is made to the above description.
  • the invention comprises a method for detecting germs in a household appliance, in particular in a washing machine or in a coffee machine.
  • This method initially involves providing a biosensor in the form of a lateral flow test strip in the above-described
  • a liquid sample or swab sample is taken and contacted with the lateral flow test strip.
  • the lateral flow test strip for example, directly into a liquid, for example, in a water tank, the household appliance, immersed.
  • the implementation of the lateral flow Tests and the detection of any present in the sample germs according to the immunological test principle based on capillary forces described above.
  • the mobility properties of the germ-specific antibodies of the test strip are used, which change depending on a complexation with the germ to be detected.
  • a readout of the lateral flow test strip takes place.
  • an optical read-out by the user takes place, a colored detection system preferably being provided for the test strip.
  • the biosensor according to the invention provides a uniform and at the same time simple technical platform in the form of the lateral flow test strip, which can be obtained by the use of different immunological detection reagents and
  • the design as a lateral flow test strip which is preferably designed as a disposable article, a very cost-effective and suitable for everyday use execution is possible.
  • Sampling point or a plug-in cartridge an automated test can be realized.
  • Coffee maker can be set up so that e.g. a sample is taken at regular intervals in an automated manner and applied to a test strip for germ analysis.
  • the evaluation and evaluation of the test strip can optionally be carried out automatically and the result can be displayed, for example, on a display, so that necessary and suitable cleaning measures or the like can be carried out.
  • FIG. 1 shows a schematic view of a preferred embodiment of a lateral flow test strip as a biosensor according to the invention.
  • Fig. 2 are schematic representations (A, B, C) of a test strip for
  • FIG. 1 shows a biosensor in the form of a lateral flow test strip 10 in a schematic view.
  • the test strip 10 is provided for the immunological detection of germs 50, depending on the germ to be detected, germ-specific antibodies 40 are present.
  • a detection reagent 41 is preferably bound directly to each antibody 40, wherein the detection reagents 41 are, for example, metal particles or
  • the lateral flow test strip 10 can be used to detect germs, in particular of molds, mold spores or bacteria, in household appliances, for example in washing machines or in coffee machines.
  • the biosensor according to the invention can generally be used for checking a germ load in all devices or devices which have, for example, permanently moist spots or other sites prone to germs.
  • the biosensor according to the invention can in principle also be used generally for a germ test in a liquid or solid sample.
  • the test strip 10 comprises a sample-receiving pad 11, a capillary 12 and a detection field 13 and a control field 14.
  • the germ-specific antibody 40 together with the detection reagent 41 bound thereto can be provided, for example, in dried form in the area of the sample-receiving pad 11.
  • the antibody 40 may be transferred to the test strip along with a buffer applied to the test strip 10, such as in the region of the sample receiving pad 11, before or after sample application.
  • the user first of all receives a sample, for example from a washing machine (eg from the drum or a washing powder compartment) or from a coffee machine, for example from the water tank, the pomace or from the region of the grinder or Brewing group, taken. This may be a liquid sample or a swab specimen.
  • Swab sample can be used, for example, a swab.
  • the sample is then applied to the sample receiving pad 11.
  • the test strip 10 with the region of the sample-receiving pad 11 is immersed directly in the liquid sample.
  • the test strip 10 may have an extension in the form of an attached rod or the like.
  • a buffer cartridge 30 may be provided, from which the buffer is released manually if necessary or optionally in an automated form.
  • buffering is generally not mandatory.
  • the germs 50 bind specifically to the antibodies 40 (antigen-antibody binding), so that complexes are formed. These complexes have changed their running properties compared to the free antibodies 40, resulting in a separation in the capillary 12.
  • the free antibody 40 runs to a certain extent further or faster than the complex of antibody 40 and germ 50.
  • the free antibody 40 reaches the remoter control field 14 further away.
  • the complex of antibody 40 and mold spore 50 reaches "only" the detection field 13. This running behavior is through a
  • Detection reagent 41 for example, a discoloration due to the metal particles in the detection field 13 and / or in the control panel 14.
  • a direct or indirect color reaction can be made in the detection field 13 and / or in the control panel 14, which can be determined by simple optical inspection of the Test strip 10 can be evaluated.
  • the color reaction can be read by the user whether the germ was in the sample or not.
  • the user receives a qualitative proof or a semiquantitative or quantitative result, the equipment of the test strip 10 and in particular the
  • Concentration of the reagents, in particular the antibody 40 is preferably designed so that a threshold for a problematic concentration of the detected germ is taken into account.
  • an optical device for example a camera, can be used for the evaluation.
  • the antibody is bound directly or directly to a dye or metal particles, so that the presence of the antibody in the respective readout field (detection field and / or control field) is directly recognizable as a color reaction.
  • the respective readout field detection field and / or control field
  • detection reagents Interaction with other detection reagents takes place, for example by known per se enzymatic detection reactions.
  • corresponding detection reagents in particular in the area of the detection field and optionally the control field, must be integrated in the test strips.
  • the control panel 14 is in particular designed so that when correct
  • test strip 10 in each case a reaction in the control field is detectable, wherein in the control panel, for example, in the above
  • FIG. 2 shows with reference to the representations A, B and C of an inventive
  • Test Strip 200 shows the possible results when performing a
  • a user can easily independently check whether, for example, a germ load is present in a washing machine or a coffee machine.
  • the user Before a cleaning action to be carried out on a regular basis, the user can check, for example, whether a germ load is actually present and a cleaning is required. In this way, for example, the

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Urology & Nephrology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Hematology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Physics & Mathematics (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un biocapteur destiné à la détection de germes (50) par test immunologique. Le biocapteur est réalisé sous la forme d'une bandelette réactive (10) pour test immunochromatographique, ladite bandelette réactive (10) comportant des anticorps (40) spécifiques d'un germe. Les propriétés de mobilité des anticorps (40) varient lorsque ces anticorps (40) forment un complexe avec le germe à détecter (50), cette variation des propriétés de mobilité étant utilisée pour la détection des germes (50).
EP16767176.7A 2015-09-25 2016-09-07 Biocapteur et procédé de détection de germes Withdrawn EP3353549A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102015218442.0A DE102015218442A1 (de) 2015-09-25 2015-09-25 Biosensor und Verfahren zum Nachweis von Keimen
PCT/EP2016/071040 WO2017050571A1 (fr) 2015-09-25 2016-09-07 Biocapteur et procédé de détection de germes

Publications (1)

Publication Number Publication Date
EP3353549A1 true EP3353549A1 (fr) 2018-08-01

Family

ID=56958888

Family Applications (1)

Application Number Title Priority Date Filing Date
EP16767176.7A Withdrawn EP3353549A1 (fr) 2015-09-25 2016-09-07 Biocapteur et procédé de détection de germes

Country Status (5)

Country Link
EP (1) EP3353549A1 (fr)
JP (1) JP2018533002A (fr)
CN (1) CN108027370A (fr)
DE (1) DE102015218442A1 (fr)
WO (1) WO2017050571A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110806477A (zh) * 2018-08-06 2020-02-18 国家纳米科学中心 一种病原菌检测试纸条、传感器及其应用
JP7350268B2 (ja) * 2019-05-30 2023-09-26 国立大学法人北海道大学 物質検出装置
WO2021193750A1 (fr) * 2020-03-26 2021-09-30 住友化学株式会社 Anticorps monoclonal, méthode de détection de bactéries, kit de détection de bactéries et hybridome
WO2023170544A1 (fr) * 2022-03-07 2023-09-14 Das Tanisha Dispositif analytique en papier (pad) pour la détection d'infections vaginales

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19604195C1 (de) 1996-02-06 1997-04-17 Hoechst Ag Verfahren zur Herstellung von Phosphonigsäuremonoalkylestern
EP1972938B1 (fr) * 2007-03-19 2014-05-14 Ivoclar Vivadent Bande de test pour la détermination du risque de carie
US20130196310A1 (en) * 2008-05-20 2013-08-01 Rapid Pathogen Screening, Inc. Method and Device for Combined Detection of Viral and Bacterial Infections
EP2288916B1 (fr) * 2008-06-17 2015-04-22 Erber Aktiengesellschaft Procédé d'immunochromatographie et système de test pour la détermination d'au moins un analyte dans une solution test à analyser
DE102010064251A1 (de) 2010-12-28 2012-06-28 Robert Bosch Gmbh Handmessgerät zum Nachweis von verdeckten Schimmelschäden in Innenräumen
EP2705366B1 (fr) * 2011-05-04 2017-07-12 Pop Test LLC Dispositif de diagnostic
AU2014226175A1 (en) * 2013-03-07 2015-07-23 Rapid Pathogen Screening, Inc. Multiplanar lateral flow assay with diverting zone

Also Published As

Publication number Publication date
DE102015218442A1 (de) 2017-03-30
WO2017050571A1 (fr) 2017-03-30
JP2018533002A (ja) 2018-11-08
CN108027370A (zh) 2018-05-11

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